AKT1
Reactivity: Human
WB, IHC, IF, IP
Host: Rabbit
Monoclonal
unconjugated
Application Notes
ELISA: Use at 0.05 μg/mL. Immunoblotting (Western): 0.5 μg/mL for HRPO/ECL detection. Recommended buffer: Casein/Tween 20 based blocking and blot incubation buffer. Provided Positive Control: Cell lysate from pervanadate treated HepG2 cells. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
Positive Control: Cell lysate from HepG2 pervanadate treatedFormat: Lyophilized cell lysate from HepG2 cells. Serum starved cells were treated for 15min. with pervanadateReconstitution: Restore by addition of 200 µl H2O. After complete solubilization add 200µl 2x SDS-PAGE sample buffer, mix and incubate at 90°C for 5 min. Application: The positive control cell lysate is recommended for immunoblot applications. 20 µl of positive control cell lysate correspond to ca. 20. 000 cells. Use 20 µl/lane (mini gel) for HRPO/ECL detection of the target proteins. Please NOTE: The lyophilized cell lysates conatin SDS and are not recommended forapplications with native proteins such as in immunoprecipitation.
Restrictions
For Research Use only
Reconstitution
Restore with 1 mL H2O (15 min, RT).
Buffer
1 mL2 x PBS / 0.09 % Sodium Azide / PEG and Sucrose.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
-20 °C
Storage Comment
For long-term storage, freeze lyophilizate upon arrival (-20 °C). Upon reconstitution,aliquote and freeze in liquid nitrogen, reconstituted antibody can be stored frozen at -80 °Cup to 1 year. Thaw aliquots at 37 °C. Thawed aliquots may be stored at 4 °C up to 3 months. Avoid repeated freeze / thaw cycles.
The PKB (RAC/akt) protein kinases are a family of second messenger-regulated serine/threonine kinases. Three mammalian isoforms (alpha, beta, gamma) have been identified so far, the alpha-isoform being the cellular homologue of the v-akt oncogene. Stimulation of cells with growth factors leads to activation of PKB by a phosphoinositide-3-kinase dependent signal transduction pathway. Activated PKB is phosphorylated at serine 473 (mTOR/Rictor or DNA-PK) and threonine 308 (PDK1).Synonyms: Akt-1, C-AKT, Protein kinase B, RAC-PK-alpha, RAC-alpha serine/threonine-protein kinase